Immunocytochemical evaluation of reorganisation of keratinocyte cytoskeleton induced by change in Ca 2+ concentration in culture medium

Abstract

Ca²⁺ level-induced changes in the arrangement of cytoskeleton of cultured keratinocytes were estimated immunocytochemically, by evaluating expression of specific cytokeratins, desmoplakin and tubulin. Keratinocytes were isolated from fragments of skin of dead human foetuses. Culture of epidermal cells was performed in two phases: phase I yielded cells of high proliferation abilities in serumfree Keratinocyte SFM of low Ca²⁺ level (0.03 mM); in phase II differentiated cells were obtained in Dulbecco medium of a high Ca2+ concentration (1.2 mM). Immunocytochemical evaluation of phase I and II cells revealed an array of differences which involved mainly expression and distribution of specific cytokeratins, distribution of tubulin, testifying to a different microtubule arrangement and distribution of desmoplakin and indicating a tendency to form desmosomes. The changes were induced by the changes in Ca²⁺ level in the culture medium.